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. 2019 Nov 11;13(1):332–344. doi: 10.1080/19336918.2019.1685928

Figure 4.

Figure 4.

Secretory AGR2 helps to organize fibroblasts by promoting cell elongation and proliferation. (a) Effect of CM collected after 5 days from MCF7, H460, SKOV3 cultures on the growth of 3T3 cells, observed after 24 h using phase contrast microscopy at x100 magnification. (b) The proliferation of NIH3T3 cells plotted after 24 h by counting cells using haemocytometer. (c) The expression of AGR2 analysed from MCF7, H460 and SKOV3 cell lysate and conditioned medium by western blot and immunoprecipitation as indicated. 50 ml conditioned medium of each culture was used for immunoprecipitation. (d) Effect of rAGR2, bFGF, and rAGR2-bFGF on the growth of 3T3 observed using phase contrast microscopy at x100 magnification. The concentration of rAGR2 is 500 ng/ml. The concentration of bFGF is 1 ng/ml. (e) The proliferation of 3T3 plotted after 24 h by counting cells using haemocytometer. Each column represents mean±SD (n = 5). (f) Cells were seeded and treated with MCF7 CM, 18A4 mAb and MCF7 CM/AGR2¯ and typical morphology were obtained by phase contrast microscopy. Scale bar: 100 µm (g) Ratio (length/width) was obtained using Image-Pro Plus software by using 80 isolated cells for each condition. Asterisks indicate the significant difference of cells proliferation and elongation. (**p < 0.001, ***p < 0.0001).